Zhang Chang-Hua, He Yu-Long, Wang Fang-Jin, Song Wu, Yuan Xi-Yu, Yang Dong-Jie, Chen Chuang-Qi, Cai Shi-Rong, Zhan Wen-Hua
Department of Gastrointestinopancreatic Surgery of the First Affiliated Hospital, Gastric Cancer Center, Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China.
World J Gastroenterol. 2008 Jan 14;14(2):298-302. doi: 10.3748/wjg.14.298.
To establish and validate the mutation testing for identification and characterization of hereditary non-polyposis colorectal cancer (HNPCC) in suspected Chinese patients.
Five independent Chinese kindreds with HNPCC fulfilling the classical Amsterdam criteria were collected. Genomic DNA was extracted after informed consent was obtained. The coding region of hMSH2 and hMLH1 genes was detected by polymerase chain reaction (PCR) and denaturing high-performance liquid chromatography (DHPLC). Mutations identified in the proband by DHPLC were directly sequenced using a 377 DNA sequencer, analyzed with a basic local alignment tool (BLAST), and tested in the corresponding family members by direct DNA sequencing.
Mutations were identified in two Chinese HNPCC kindreds. One was the missense mutation of hMSH2 c.1808A-->G resulting in Asp 603 Gly identified in the proband of the fifth HNPCC (HNPCC5) kindred. In the HNP5 kindred, three family members were found to have this mutation and two of them had colorectal cancer. The other mutation of hMLH1 c.1882A-->G was identified in the HNP2 kindred's proband, which might be the nonsense mutation analyzed by BLAST.
Pedigree investigation and mutation testing of hMSH2 and hMLH1 are the practical methods to identify high-risk HNPCC patients in China.
建立并验证用于鉴定和表征疑似中国患者遗传性非息肉病性结直肠癌（HNPCC）的突变检测方法。
收集了五个符合经典阿姆斯特丹标准的中国HNPCC独立家系。在获得知情同意后提取基因组DNA。通过聚合酶链反应（PCR）和变性高效液相色谱（DHPLC）检测hMSH2和hMLH1基因的编码区。通过DHPLC在先证者中鉴定出的突变，使用377 DNA测序仪进行直接测序，用基本局部比对工具（BLAST）进行分析，并通过直接DNA测序在相应的家庭成员中进行检测。
在两个中国HNPCC家系中鉴定出突变。一个是在第五个HNPCC（HNPCC5）家系的先证者中鉴定出的hMSH2 c.1808A→G错义突变，导致Asp 603 Gly。在HNP5家系中，发现三名家庭成员有此突变，其中两名患有结直肠癌。另一个hMLH1 c.1882A→G突变在HNP2家系的先证者中被鉴定出来，这可能是经BLAST分析的无义突变。
家系调查以及hMSH2和hMLH1的突变检测是在中国识别高危HNPCC患者的实用方法。
